State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
Appl Microbiol Biotechnol. 2010 Jun;87(1):281-7. doi: 10.1007/s00253-010-2457-y. Epub 2010 Feb 16.
Pseudomonas sp. strain NyZ402, a native soil organism that grows on para-nitrophenol (PNP), was genetically engineered for the simultaneous degradation of methyl parathion (MP) and ortho-nitrophenol (ONP) by integrating mph (methyl parathion hydrolase gene) from Pseudomonas sp. strain WBC-3 and onpAB (ONP 2-monooxygenase and ONP o-benzoquinone reductase genes) from Alcaligenes sp. strain NyZ215 into the genome of strain NyZ402. Methyl parathion hydrolase (MPH), ONP 2-monooxygenase (OnpA) and o-benzoquinone reductase (OnpB) were constitutively expressed in the engineered strain NyZ-MO. Strain NyZ-MO was free of exogenous antibiotic resistance gene markers and the introduced genes were genetically stable. Degradation experiments showed that strain NyZ-MO could utilize MP or ONP as the sole carbon and energy source, and mineralize 0.1 mM MP-0.1 mM ONP simultaneously. This method may serve as a useful strategy for the construction of engineered strains in the degradation of multiple environmental pollutants.
假单胞菌（Pseudomonas sp.）菌株 NyZ402 是一种天然土壤微生物，能够以对硝基苯酚（PNP）作为唯一碳源和能源生长，现已通过基因工程手段，将甲基对硫磷水解酶基因（mph）和邻硝基苯酚 2-单加氧酶基因（onpAB）整合到菌株 NyZ402 的基因组中，实现了对甲基对硫磷和邻硝基苯酚的同时降解。该工程菌株 NyZ-MO 中，甲基对硫磷水解酶（MPH）、邻硝基苯酚 2-单加氧酶（OnpA）和邻硝基苯酚 o-苯醌还原酶（OnpB）均为组成型表达。工程菌株不含外源抗生素抗性基因标记，且导入基因遗传稳定。降解实验表明，菌株 NyZ-MO 可以将 MP 或 ONP 作为唯一碳源和能源利用，并同时矿化 0.1 mM MP 和 0.1 mM ONP。该方法可能为构建同时降解多种环境污染物的工程菌提供了一种有用的策略。
